Download Job Description - Faculty of Biological Sciences

Faculty of Biological Sciences
School of School of Molecular and Cellular Biology
Summer Research Placement
8 weeks, between July – September 2016
Job Summary
The Picornaviridae family of viruses contain many important pathogens, such as
poliovirus and hepatitis A virus which cause devastating diseases in mammals.
Members of the Picornavirales have a single stranded, positive sense RNA genome
encapsidated in a non-enveloped capsid with pseudo T=3 icosahedral symmetry.
Despite detailed structural information for the capsid of Picornavirales, the
mechanism for genome encapsidation is not understood. Cowpea mosaic virus
(CPMV) is a plant-infecting member of the Picornavirales which is of major interest in
biotechnology, and is an ideal model system to study genome encapsidation. A
better understanding of CPMV genome encapsidation will allow us to further develop
CPMV biotechnology applications. We aim to use high-resolution cryo-electron
microscopy (EM) analysis of CPMV and CPMV mutants to unravel the mechanism of
genome packaging. We have produced an atomic resolution cryo-EM structure of
wild-type (WT) CPMV and an empty virus-like (eVLP). Using the information from
both eVLP and CPMV maps we have designed, produced and purified mutant CPMV
particles that are unable to package RNA. A F192W mutant, contains a conservative
mutation of an amino acid located on the external surface of CPMV, which is unable
to encapsidate RNA but can produce viral particles.
You will analyse the structure of CPMV F192W using cryo-EM. You will make and
assist in screening cryo-EM grids of CPMV F192W to ensure high quality grids are
produced using our current 200 kV electron microscope. Optimal grids will be
analysed using our new 300 kV Titan Krios electron microscope (due to be delivered
May 2016). We will collect a large cryo-EM data set for the CPMV F192W mutant.
We anticipate collecting ~1,500 micrographs using a direct electron detector. We aim
to produce high-resolution reconstructions of the CPMV mutant using single particle
image processing software (RELION) to produce a 3D map. Using this method we
have already produced high-resolution WT CPMV and an CPMV empty virus like
particles (eVLP) maps (3.4 Å and 3.0 Å respectively) (Hesketh et al., Nat. Comms,
2015). We will compare the structure of WT CPMV to the structure of the mutant
CPMV to gain further insights into CPMV biogenesis. Using this information we aim
to understand better how CPMV recognises it’s RNA genome for encapsidation prior
to capsid assembly.
Stipend is £200 per week.
Informal enquiries may be made to Emma Hesketh, email:
[email protected].
To apply, please send your CV and a covering letter (200-300 words maximum)
Closing Date: Noon on 9th March 2016
Interviews: 16th March
Job Description
Reports to: Emma Hesketh
Main Duties and Responsibilities

Prepare samples for negative stain and cryo-electron microscopy (cryo-EM).

Process cryo-EM data using single particle image processing software such
as RELION.

Write up technical procedures and methods into detailed protocols, to allow
reproducibility of experiments by others.

Present data in an understandable manner to members of the group.
Person Specification
Essential
 Ability to work independently after induction period, manage and prioritise own
tasks on a day-to-day basis with little supervision.
 Good organisational skills and time management skills.
 Highly motivated and hard working.
Desirable
 Knowledge of UNIX and the command line.
Additional Information
Details of the terms and conditions of employment for all staff at the university,
including information on pensions and benefits, are available on the Human
Resources web pages accessible via the links on the right hand side, or at
http://www.leeds.ac.uk/hr/index.htm
Criminal Record Disclosures
A Criminal Records Disclosure is not required for this position. However, applicants
who have unspent convictions must indicate this in the ‘other personal details’
section of the application.
Disabled Applicants
The post is located in the Faculty of Biological Sciences. Disabled applicants wishing
to review access to the building are invited to contact the department direct.
Additional information may be sought from the Recruitment Officer, email
[email protected] or tel + 44 (0)113 343 1723.
Disabled applicants are not obliged to inform employers of their disability but will still
be covered by the Disability Discrimination Act once their disability becomes known.
Further information for applicants with disabilities, impairments or health
conditions is available in the applicant guidance.
University Values
All staff are expected to operate in line with the university’s values and standards,
which work as an integral part of our strategy and set out the principles of how we
work together. More information about the university’s strategy and values is
available at http://www.leeds.ac.uk/comms/strategy/